DOCSLIB.ORG

Analysis of the Metaphase Chromosome Karyotypes in Imaginal Discs of Aedes Communis, Ae. Punctor, Ae. Intrudens, and Ae. Rossicus (Diptera: Culicidae) Mosquitoes

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Analysis of the Metaphase Chromosome Karyotypes in Imaginal Discs of Aedes Communis, Ae. Punctor, Ae. Intrudens, and Ae. Rossicus (Diptera: Culicidae) Mosquitoes insects Article Analysis of the Metaphase Chromosome Karyotypes in Imaginal Discs of Aedes communis, Ae. punctor, Ae. intrudens, and Ae. rossicus (Diptera: Culicidae) Mosquitoes Svetlana S. Alekseeva 1,2,* , Yulia V. Andreeva 1, Irina E. Wasserlauf 1, Anuarbek K. Sibataev 1 and Vladimir N. Stegniy 1 1 Laboratory of Evolution Cytogenetics, Tomsk State University, Lenin st, 36, Tomsk 634050, Russia; [email protected] (Y.V.A.); [email protected] (I.E.W.); [email protected] (A.K.S.); [email protected] (V.N.S.) 2 Laboratory of Evolutionary Genomics of Insects, the Federal Research Center Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of Sciences, Lavrentiev ave., 10, Novosibirsk 630090, Russia * Correspondence: [email protected]; Tel.: +7-913-889-92-89 Received: 11 December 2019; Accepted: 16 January 2020; Published: 19 January 2020 Abstract: In this study, cytogenetic analysis of the metaphase chromosomes from imaginal discs of Aedes (Diptera: Culicidae) mosquitoes—Aedes communis, Ae. punctor, Ae. intrudens, and Ae. rossicus—was performed. The patterns of C-banding and DAPI staining of the heteroсhromatin and the length of the chromosomes demonstrate species specificity. In particular, the Ae. punctor chromosomes are the shortest compared with Ae. communis, Ae. intrudens, and Ae. rossicus, and they also carry additional C and DAPI bands in intercalary regions. The Ae. intrudens chromosomes are the longest, they have pericentromeric C bands, and they almost lack any DAPI bands near the centromere of chromosome 3 versus Ae. communis, which has the largest pericentromeric DAPI blocks in all three chromosome pairs. Ae. rossicus also possesses DAPI bands in the centromeric regions of all chromosomes, but their staining is weaker compared with those of Ae. communis. Therefore, the analysis of karyotypes is a tool for species-level identification of these mosquitoes. Keywords: mosquitoes; Culicidae; Aedes; C-banding; DAPI; mitotic chromosomes; imaginal discs 1. Introduction The Aedes mosquitoes are vectors of various diseases, determining the relevance of corresponding studies. In particular, West Nile virus, first isolated in 1937 in Uganda, is now more widespread and currently in Russia (Volgograd, Astrakhan, and Rostov oblasts and Krasnodar Krai). In nature, this virus is transmitted as follows: bird mosquito another vertebrate [1]. Dengue fever, widespread $ ! in regions with a tropical and subtropical climate, was discovered on Madeira Island and in several southern regions of Europe [2]. Zika virus, earlier prevalent in African and Asian countries, currently causes outbreaks in America [3]. Dirofilariasis is characteristic of regions with a humid and hot climate; however, an increase in dirofilariasis morbidity has recently been observed in countries for which it was previously rather untypical [4,5]. Ae. communis, Ae. punctor, Ae. cinereus, Culex pipiens, and some other species activity cycles have been studied in Sweden because of the transmission of Ockelbo disease (caused by Sindbis virus) and tularaemia in Sweden [6]. Aedes mosquitoes are vectors of these and many other diseases [2,7–10]. Mosquitoes from the genus Aedes have been distributed all over the world from their original habitat. Currently, invasive species of Aedes mosquitoes—Ae. albopictus, Ae. japonicus, Ae. atropalpus, Insects 2020, 11, 63; doi:10.3390/insects11010063 www.mdpi.com/journal/insects Insects 2020, 11, 63 2 of 9 Ae. koreicus, and Ae. aegypti [11–19]—are being ever more frequently discovered in Europe and other regions of the world. Ae. rossicus was found close to the Arctic circle in northern Sweden [20]. Therefore, prediction of the epidemiological threat and its control requires knowledge about the species composition of the corresponding mosquito vectors. A more precise species-level identification of mosquitoes requires a set of different methods. Morphological and molecular methods are sometimes not enough for some species of mosquitoes from the Aedes genus. This is a reason to perform a karyotype analysis for Aedes mosquitoes. As is known, the amount and distribution pattern of heterochromatin in chromosomes is among the species-specific characteristics for most plants and animals [21]; thus, heterochromatin is an important object in genomic studies of individual organisms [22]. For example, a comparative karyotype analysis of the flies Lucilia cluvia and L. sericata detected differences in the chromatin structure by C-banding and other staining types [23]. A karyotype analysis of four species—Triceratopyga calliphoroides, L. porphyrina, Chrysomya pinguis, and Xenocalliphora hortona—demonstrated differences in sex chromosomes and similarity in autosomes [24]. A study of the amount and distribution of heterochromatin on the chromosomes of several groups of closely related species (complexes of Drosophila, Anopheles, and Bactrocera)[25] demonstrated that a quantitative assay of heterochromatin in mitotic chromosomes can be used for the identification of cryptic species. Differences in the heterochromatin amount and localization have been demonstrated for two sibling species, Anopheles atroparvus and An. labranchiae [26]. Previously, we performed a comparative analysis of the metaphase chromosomes in imaginal discs of the mosquito species Ae. excrucians, Ae. behningi, and Ae. euedes and showed that heterochromatin patterns of chromosomes represent one of the characteristics for the species-level identification of mosquitoes [27]. The previous karyotype analysis involved Ae. excrucians, Ae. behningi, and Ae. euedes mosquitoes collected in the Tomsk region (south of Western Siberia, Russia), which houses 21 Aedes mosquito species [28], and which are also widely abundant in other countries (http://www.mosquitocatalog.org/default.aspx). This (current) analysis includes four more species (Ae. communis, Ae. punctor, Ae. intrudens, and Ae. rossicus) from the Aedes genus in the Tomsk region (south of Western Siberia, Russia). We collected them in May–June (2019) and performed a chromosome analysis. The goal of this work was to analyze the karyotypes of Aedes (Diptera: Culicidae) mosquitoes (Ae. communis, Ae. punctor, Ae. intrudens, and Ae. rossicus) in order to find out if species-specific features exist in their metaphase chromosomes. 2. Materials and Methods The 4th instar larvae of Ae. communis, Ae. punctor, Ae. intrudens, and Ae. rossicus examined in this work were sampled in water bodies of the Tomsk region. Morphological species-level identification of the sampled larvae was conducted using MBS-12 (Russia) and Stemi 2000-C (Carl Zeiss, Germany) stereo microscopes, according to the conventional descriptions and keys [29–31]. The nomenclature is given according to the Systematic Catalog of Culicidae (http://mosquitocatalog.org/default.aspx). Larvae were fixed with Carnoy’s solution (ethanol to glacial acetic acid, 3:1). Metaphase plates of dividing imaginal disc cells of the early 4th instar larvae were examined. The structure of metaphase chromosomes was assayed using lacto-aceto-orcein staining [32], C-banding, and DAPI staining [33]. 2.1. Lacto-Aceto-Orcein Staining Imaginal discs were dissected from Ae. behningi, Ae. euedes, and Ae. excrucians larvae fixed with Carnoy’s solution, stained in a drop of lacto-aceto-orcein dye for 15 min, and washed in 45% acetic acid. The stained imaginal discs were covered with a cover glass to get squash preparations by tapping on the cover glass. The squash preparations were examined using a Zeiss Axioimager A1 (Zeiss, Jena, Germany) light microscope. Insects 2020, 11, 63 3 of 9 2.2. DAPI Staining For this purpose, imaginal discs were isolated from mosquito larvae in a drop of Carnoy’s solution, transferred to a drop of 45% acetic acid, covered with a cover glass, and squashed. The cover glass was removed using liquid nitrogen and the preparations were dehydrated by successive treatment with alcohols (50%, 70%, and 96%; 5 min each). A drop of DAPI (a fluorescent dye) was placed onto air-dried preparations, which were then covered with a cover glass. The resulting slides with DAPI-stained metaphase chromosomes were examined using a Zeiss Axioimager Z1 (Zeiss, Germany) fluorescence microscope. 2.3. C-Banding C-banding was performed using the pre-staining of chromosome preparations with Ba(ОН)2. The air-dried preparations of mosquito imaginal discs were incubated in 0.2 M HCl at room temperature for 1 h and placed in fresh 5% barium hydroxide solution at 50 ◦C for 10–15 min. Then, the preparations were washed and incubated in 2 SSC buffer at 60 C for 1 h. The resulting slides were washed, × ◦ stained with 4% Giemsa solution for 1.5 h, and examined using a Zeiss Axioimager A1 (Zeiss, Germany) microscope. 2.4. Statistical Analysis The chromosomes were identified based on the ratio of their arms and lengths, according to the relevant chromosome classification [34]. The lengths of chromosomes and their arms were measured using the ImageJ program. The centromeric index was calculated as Jc = p/(p + q), where p is the short chromosome arm and q is the long arm. The relative chromosome length was calculated as Length of chromosome Lr = 100%, Total length of all chromosomes × where Lr is the relative chromosome length (%). Over 50 metaphase plates were examined for each species and 30 metaphase plates with the same degrees of condensation were selected for analysis. The p value was calculated in the Statistica 10 program for each chromosome of each species. 3. Results Karyotype analysis of the metaphase
Load more

Recommended publications
  • Hierarchical Looping of Zigzag Nucleosome Chains in Metaphase Chromosomes
    Hierarchical looping of zigzag nucleosome chains in metaphase chromosomes Sergei A. Grigoryeva,1, Gavin Bascomb, Jenna M. Buckwaltera, Michael B. Schuberta, Christopher L. Woodcockc, and Tamar Schlickb,d,1 aDepartment of Biochemistry and Molecular Biology, Milton S. Hershey Medical Center, Pennsylvania State University College of Medicine, Hershey, PA 17033; bDepartment of Chemistry and Courant Institute of Mathematical Sciences, New York University, New York, NY 10012; cBiology Department, University of Massachusetts, Amherst, MA 01003; and dNYU-ECNU Center for Computational Chemistry, NYU Shanghai, Shanghai 200062, China Edited by Michael Levitt, Stanford University School of Medicine, Stanford, CA, and approved December 22, 2015 (received for review September 14, 2015) The architecture of higher-order chromatin in eukaryotic cell nuclei is However, evidence for 30-nm fibers in interphase nuclei of living largely unknown. Here, we use electron microscopy-assisted nucleo- cells has been controversial (reviewed in refs. 9 and 10). For exam- some interaction capture (EMANIC) cross-linking experiments in ple, whereas a distinct 30-nm fiber architecture is observed in ter- combination with mesoscale chromatin modeling of 96-nucleosome minally differentiated cells (11, 12), neither continuous nor periodic arrays to investigate the internal organization of condensed chroma- 30-nm fibers are observed in the nuclei of proliferating cells (13–15). tin in interphase cell nuclei and metaphase chromosomes at nucleo- However, zigzag features of the chromatin fibers are well supported somal resolution. The combined data suggest a novel hierarchical by nucleosome interaction mapping in vitro (16) and in vivo (15). looping model for chromatin higher-order folding, similar to rope For chromatin architecture within metaphase chromosomes, flaking used in mountain climbing and rappelling.
    [Show full text]
  • Ftsk Actively Segregates Sister Chromosomes in Escherichia Coli
    FtsK actively segregates sister chromosomes in Escherichia coli Mathieu Stoufa,b, Jean-Christophe Meilea,b, and François Corneta,b,1 aLaboratoire de Microbiologie et de Génétique Moléculaires, Centre National de la Recherche Scientifique, F-31000, Toulouse, France; and bUniversité Paul Sabatier, Université de Toulouse, F-31000, Toulouse, France Edited by Nancy E. Kleckner, Harvard University, Cambridge, MA, and approved May 23, 2013 (received for review March 6, 2013) Bacteria use the replication origin-to-terminus polarity of their cir- with the divisome, is also required (13, 14). FtsK acts in a region cular chromosomes to control DNA transactions during the cell cy- about 400 kb long (15) and translocates DNA toward dif.Trans- cle. Segregation starts by active migration of the region of origin location is oriented by recognition of the FtsK-orienting polar followed by progressive movement of the rest of the chromo- sequences (KOPS) DNA motifs that are preferentially oriented somes. The last steps of segregation have been studied extensively toward dif, particularly in the ter region (4, 16–18). Upon reaching in the case of dimeric sister chromosomes and when chromosome the dif site, FtsK activates XerCD-mediated recombination that organization is impaired by mutations. In these special cases, the resolves chromosome dimers. The oriented translocation activity divisome-associated DNA translocase FtsK is required. FtsK pumps of FtsK also is strictly required when chromosome organization is chromosomes toward the dif chromosome dimer resolution site impaired by mutations, for instance by inactivation of the MukBEF using polarity of the FtsK-orienting polar sequence (KOPS) DNA complex (19, 20) or in strains carrying important asymmetry of the motifs.
    [Show full text]
  • Organization, Evolution and Function of Alpha Satellite Dna
    ORGANIZATION, EVOLUTION AND FUNCTION OF ALPHA SATELLITE DNA AT HUMAN CENTROMERES by M. KATHARINE RUDD Submitted in partial fulfillment of the requirements For the degree of Doctor of Philosophy Dissertation Advisor: Dr. Huntington F. Willard Department of Genetics CASE WESTERN RESERVE UNIVERSITY January, 2005 CASE WESTERN RESERVE UNIVERSITY SCHOOL OF GRADUATE STUDIES We hereby approve the dissertation of ______________________________________________________ candidate for the Ph.D. degree *. (signed)_______________________________________________ (chair of the committee) ________________________________________________ ________________________________________________ ________________________________________________ ________________________________________________ ________________________________________________ (date) _______________________ *We also certify that written approval has been obtained for any proprietary material contained therein. 1 Table of Contents Table of contents.................................................................................................1 List of Tables........................................................................................................2 List of Figures......................................................................................................3 Acknowledgements.............................................................................................5 Abstract................................................................................................................6
    [Show full text]
  • Comparative Genomic Hybridization in the Detection of DNA Copy Number Abnormalities in Uveal Melanoma1
    [CANCER RESEARCH 54. 4764-4768. September 1. 1994] Comparative Genomic Hybridization in the Detection of DNA Copy Number Abnormalities in Uveal Melanoma1 Kathleen B. Gordon, Curtis T. Thompson, Devron H. Char,2 Joan M. O'Brien, Stewart Kroll, Siavash Ghazvini, and Joe W. Gray Ocular Oncology Unii IK. B. G., D. H. C., J. M. O., S. K., S. G.¡and Laboratory of Molecular Cylomelry ¡C.T. T., J. W. G.I, University of California, San Francisco, California 94143-0730 ABSTRACT identified, and the possibility that more than one locus is involved in tumor initiation and progression can be assessed. Genomic DNA from Genomic instability appears to play an important role in the develop tumor specimens is used so that genetic alterations identified with ment, growth, invasiveness, and eventual metastasis of the neoplastic cell. CGH are not artifactually altered by propagation in cell culture. In the We have used a powerful new technique, comparative genomic hybrid present study, we used CGH to detect alterations in gene copy number ization, to evaluate genetic alterations in 10 fresh frozen uveal melanomas. Comparative genomic hybridization utilizes dual fluorescence in situ hy in ten fresh frozen uveal melanomas. bridization to characterize chromosome deletions and duplications, allow ing for simultaneous evaluation of the entire human genome. Several MATERIALS AND METHODS consistent chromosomal abnormalities were detected. This study con Clinical Data. Ten uveal melanomas were evaluated after primary enucle- firmed previous findings obtained using standard cytogenetic techniques ation. The tumors were classified histologically according to the modified but demonstrated an increased incidence in abnormalities of chromo Callender classification (5).
    [Show full text]
  • Two Distinct Domains in Drosophila Melanogaster Telomeres
    Copyright Ó 2005 by the Genetics Society of America DOI: 10.1534/genetics.105.048827 Two Distinct Domains in Drosophila melanogaster Telomeres Harald Biessmann,* Sudha Prasad,† Valery F. Semeshin,‡ Eugenia N. Andreyeva,‡ Quang Nguyen,§ Marika F. Walter* and James M. Mason†,1 *Developmental Biology Center, University of California, Irvine, California 92697, †Laboratory of Molecular Genetics, National Institute of Environmental Health Sciences, Research Triangle Park, North Carolina 27709, ‡Laboratory of Molecular Cytogenetics, Institute of Cytology and Genetics, Russian Academy of Sciences, Novosibirsk 630090, Russia and §Department of Biological Chemistry, University of California, Irvine, California 92697 Manuscript received July 27, 2005 Accepted for publication August 16, 2005 ABSTRACT Telomeres are generally considered heterochromatic. On the basis of DNA composition, the telomeric region of Drosophila melanogaster contains two distinct subdomains: a subtelomeric region of repetitive DNA, termed TAS, and a terminal array of retrotransposons, which perform the elongation function instead of telomerase. We have identified several P-element insertions into this retrotransposon array and compared expression levels of transgenes with similar integrations into TAS and euchromatic regions. In contrast to insertions in TAS, which are silenced, reporter genes in the terminal HeT-A, TAHRE,orTART retroelements did not exhibit repressed expression in comparison with the same transgene construct in euchromatin. These data, in combination with cytological studies, provide evidence that the subtelomeric TAS region exhibits features resembling heterochromatin, while the terminal retrotransposon array exhibits euchromatic characteristics. NA sequences at the ends of eukaryotic chromo- tandem repeats of 457 bp (Walter et al. 1995; Mason D somes are the products of a telomere elongation et al.
    [Show full text]
  • Staining, and in Situ Digestion with Restriction Endonucleases
    Heredity66 (1991) 403—409 Received 23 August 1990 Genetical Society of Great Britain An analysis of coho salmon chromatin by means of C-banding, AG- and fluorochrome staining, and in situ digestion with restriction endonucleases R. LOZANO, C. RUIZ REJON* & M. RUIZ REJON* Departamento de Biologia Animal, Ecologia y Genética. E. /ngenierIa T. AgrIcola, Campus Universitario de Almeria, 04120 AlmerIa and *Facu/tad de Ciencias, 18071 Granada, Universidad de Granada, Spain Thechromosome complement of the coho salmon (Oncorhynchus kisutch) has been analysed by means of C-banding, silver and fluorochrome staining, and in situ digestion with restriction endo- nucleases. C-banding shows heterochromatic regions in the centromeres of most chromosomes but not in the telomeric areas. The fifteenth metacentric chromosome pair contains a large block of constitutive heterochromatin, which occupies almost all of one chromosome arm. This region is also the site where the ribosomal cistrons are located and it reacts positively to CMA3/DA fluorochrome staining. The NORs are subject to chromosome polymorphism, which might be explicable in terms of an amplification of ribosomal cistrons. The digestion banding patterns produced by four types of restriction endonucleases on the euchromatic and heterochromatic regions are described. Two kinds of highly repetitive DNAs can be distinguished and the role of restriction endonucleases as a valuable tool in chromosome characterization studies, as well as in the analysis of the structure and organization of fish chromatin, are also discussed. Keywords:C-banding,coho salmon, fluorochrome staining, restriction endonuclease banding. (Oncorhynchus kisutch), as well as applying conven- Introduction tional banding techniques, we have analysed the Theuse of restriction endonucleases (REs) is becom- mitotic chromosomes using DNA base-pair-specific ing common not only in molecular biology but also as fluorochromes and in situ digestion with restriction an important tool in molecular cytogenetics.
    [Show full text]
  • Holocentric Chromosomes: Convergent Evolution, Meiotic Adaptations, and Genomic Analysis
    Chromosome Res DOI 10.1007/s10577-012-9292-1 Holocentric chromosomes: convergent evolution, meiotic adaptations, and genomic analysis Daniël P. Melters & Leocadia V. Paliulis & Ian F. Korf & Simon W. L. Chan # Springer Science+Business Media B.V. 2012 Abstract In most eukaryotes, the kinetochore protein trait has arisen at least 13 independent times (four times in complex assembles at a single locus termed the centro- plants and at least nine times in animals). Holocentric mere to attach chromosomes to spindle microtubules. chromosomes have inherent problems in meiosis because Holocentric chromosomes have the unusual property of bivalents can attach to spindles in a random fashion. attaching to spindle microtubules along their entire Interestingly, there are several solutions that have evolved length. Our mechanistic understanding of holocentric to allow accurate meiotic segregation of holocentric chro- chromosome function is derived largely from studies in mosomes. Lastly, we describe how extensive genome the nematode Caenorhabditis elegans, but holocentric sequencing and experiments in nonmodel organisms chromosomes are found over a broad range of animal may allow holocentric chromosomes to shed light on and plant species. In this review, we describe how hol- general principles of chromosome segregation. ocentricity may be identified through cytological and molecular methods. By surveying the diversity of organ- Keywords centromere . holocentric . meiosis . isms with holocentric chromosomes, we estimate that the phylogeny. tandem repeat . chromosome Abbreviations Responsible Editor: Rachel O’Neill and Beth Sullivan. ChIP-seq Chromatin immunoprecipitation Electronic supplementary material The online version of this followed by sequencing article (doi:10.1007/s10577-012-9292-1) contains ChIP-chip Chromatin immunoprecipitation supplementary material, which is available to authorized users.
    [Show full text]
  • Subtelomere Organization in the Genome of the Microsporidian Encephalitozoon Cuniculi: Patterns of Repeated Sequences and Physic
    Dia et al. BMC Genomics (2016) 17:34 DOI 10.1186/s12864-015-1920-7 RESEARCH ARTICLE Open Access Subtelomere organization in the genome of the microsporidian Encephalitozoon cuniculi: patterns of repeated sequences and physicochemical signatures Ndongo Dia1*, Laurence Lavie2, Ngor Faye3, Guy Méténier2, Edouard Yeramian4, Christophe Duroure5, Bhen S. Toguebaye3, Roger Frutos6, Mbayame N. Niang1, Christian P. Vivarès2, Choukri Ben Mamoun7 and Emmanuel Cornillot8,9* Abstract Background: The microsporidian Encephalitozoon cuniculi is an obligate intracellular eukaryotic pathogen with a small nuclear genome (2.9 Mbp) consisting of 11 chromosomes. Although each chromosome end is known to contain a single rDNA unit, the incomplete assembly of subtelomeric regions following sequencing of the genome identified only 3 of the 22 expected rDNA units. While chromosome end assembly remains a difficult process in most eukaryotic genomes, it is of significant importance for pathogens because these regions encode factors important for virulence and host evasion. Results: Here we report the first complete assembly of E. cuniculi chromosome ends, and describe a novel mosaic structure of segmental duplications (EXT repeats) in these regions. EXT repeats range in size between 3.5 and 23.8 kbp and contain four multigene families encoding membrane associated proteins. Twenty-one recombination sites were identified in the sub-terminal region of E. cuniculi chromosomes. Our analysis suggests that these sites contribute to the diversity of chromosome ends organization through Double Strand Break repair mechanisms. The region containing EXT repeats at chromosome extremities can be differentiated based on gene composition, GC content, recombination sites density and chromosome landscape. Conclusion: Together this study provides the complete structure of the chromosome ends of E.
    [Show full text]
  • Genome-Wide Characterization of Satellite DNA Arrays in a Complex Plant Genome Using Nanopore Reads
    bioRxiv preprint doi: https://doi.org/10.1101/677575; this version posted June 25, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. Genome-wide characterization of satellite DNA arrays in a complex plant genome using nanopore reads by Tihana Vondrak1,2, Laura Ávila Robledillo1,2, Petr Novák1, Andrea Koblížková1, Pavel Neumann1 and Jiří Macas1,* (1) Biology Centre, Czech Academy of Sciences, Branišovská 31, České Budějovice, CZ-37005, Czech Republic (2) University of South Bohemia, Faculty of Science, České Budějovice, Czech Republic * Corresponding author e-mail: [email protected] phone: +420 387775516 1 bioRxiv preprint doi: https://doi.org/10.1101/677575; this version posted June 25, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. 1 Abstract 2 Background: Amplification of monomer sequences into long contiguous arrays is the main 3 feature distinguishing satellite DNA from other tandem repeats, yet it is also the main obstacle in 4 its investigation because these arrays are in principle difficult to assemble. Here we explore an 5 alternative, assembly-free approach that utilizes ultra-long Oxford Nanopore reads to infer the 6 length distribution of satellite repeat arrays, their association with other repeats and the 7 prevailing sequence periodicities.
    [Show full text]
  • First Chromosome Analysis and Localization of the Nucleolar Organizer Region of Land Snail, Sarika Resplendens (Stylommatophora, Ariophantidae) in Thailand
    © 2013 The Japan Mendel Society Cytologia 78(3): 213–222 First Chromosome Analysis and Localization of the Nucleolar Organizer Region of Land Snail, Sarika resplendens (Stylommatophora, Ariophantidae) in Thailand Wilailuk Khrueanet1, Weerayuth Supiwong2, Chanidaporn Tumpeesuwan3, Sakboworn Tumpeesuwan3, Krit Pinthong4, and Alongklod Tanomtong2* 1 School of Science and Technology, Khon Kaen University, Nong Khai Campus, Muang, Nong Khai 43000, Thailand 2 Applied Taxonomic Research Center (ATRC), Department of Biology, Faculty of Science, Khon Kaen University, Muang, Khon Kaen 40002, Thailand 3 Department of Biology, Faculty of Science, Mahasarakham University, Kantarawichai, Maha Sarakham 44150, Thailand 4 Biology Program, Faculty of Science and Technology, Surindra Rajabhat University, Muang, Surin 32000, Thailand Received July 23, 2012; accepted February 25, 2013 Summary We report the first chromosome analysis and localization of the nucleolar organizer re- gion of the land snail Sarika resplendens (Philippi 1846) in Thailand. The mitotic and meiotic chro- mosome preparations were carried out by directly taking samples from the ovotestis. Conventional and Ag-NOR staining techniques were applied to stain the chromosomes. The results showed that the diploid chromosome number of S. resplendens is 2n=66 and the fundamental number (NF) is 132. The karyotype has the presence of six large metacentric, two large submetacentric, 26 medium metacentric, and 32 small metacentric chromosomes. After using the Ag-NOR banding technique, one pair of nucleolar organizer regions (NORs) was observed on the long arm subtelomeric region of chromosome pair 11. We found that during metaphase I, the homologous chromosomes show synapsis, which can be defined as the formation of 33 ring bivalents, and 33 haploid chromosomes at metaphase II as diploid species.
    [Show full text]
  • Transposon Telomeres Are Widely Distributed in the Drosophila Genus: TART Elements in the Virilis Group
    Transposon telomeres are widely distributed in the Drosophila genus: TART elements in the virilis group Elena Casacuberta and Mary-Lou Pardue* Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02139 Contributed by Mary-Lou Pardue, January 20, 2003 Telomeres of most animals, plants, and unicellular eukaryotes are long stretches of noncoding sequence, which can differ signifi- made up of tandem arrays of repeated DNA sequences produced cantly between copies of the element in the same Drosophila by the enzyme telomerase. Drosophila melanogaster has an un- stock. Both elements also encode Gag proteins whose sequence usual variation on this theme; telomeres consist of tandem arrays can vary as much as that of the noncoding regions (4). TART has of sequences produced by successive transpositions of two non- a second ORF (ORF2) encoding a protein with endonuclease LTR retrotransposons, HeT-A and TART. To explore the phyloge- and reverse transcriptase (RT) activities. As with retroviruses netic distribution of these variant telomeres, we have looked for (6), ORF2 is more conserved than the Gag coding sequence, and TART homologues in a distantly related Drosophila species, virilis. the most conserved part, RT, has been the only useful probe for We have found elements that, despite many differences in nucle- these cross-species homology searches. Although the high otide sequence, retain significant amino acid similarity to TART sequence divergence makes it difficult to study the telomere from D. melanogaster. These D. virilis TART elements have features elements, it also increases the probability that conserved features that characterize TART elements in D. melanogaster:(i) they are are of biological importance.
    [Show full text]
  • Satellite DNA Sequences in the Human Acrocentric Chromosomes: Information from Translocations and Heteromorphisms
    Am J Hum Genet 33:243-251, 1981 Satellite DNA Sequences in the Human Acrocentric Chromosomes: Information from Translocations and Heteromorphisms J. R. GOSDEN,1 S. S. LAWRIE, AND C. M. GOSDEN SUMMARY Satellite III DNA has been located by in situ hybridization in chromo- somes 1, 3-5,7,9, 10, 13-18,20-22, and Y and ribosomal DNA (rDNA) in the acrocentric chromosomes 13-15, 21, and 22. In the acrocentric chro- mosomes, the satellite DNA is located in the short arm. Here we report comparisons by in situ hybridization of the amount of satellite DNA in Robertsonian translocation and "normal variant" chro- mosomes with that in their homologs. In almost all dicentric Robertsonian translocations, the amount ofsatellite DNA is less than that in the normal homologs, but it is rarely completely absent, indicating that satellite DNA is located between the centromere and the nucleolus organizer region (NOR) and that the breakpoints are within the satellite DNA. The amount of satellite DNA shows a range of variation in "normal" chromosomes, and this is still more extreme in "normal variant" chromo- somes, those with large short arms (p+ or ph+) generally having more satellite DNA than those with small short arms (p- or ph-). The cytologi- cal satellites are heterogeneous in DNA content; some contain satellite DNA, others apparently do not, and the satellite DNA content is not related to the size or intensity of fluorescence of the satellites. The signifi- cance of these variations for the putative functions of satellite DNA is discussed. INTRODUCTION The human acrocentric chromosomes (D group: chromosomes 13, 14, and 15, and G group: chromosomes 21 and 22) have been the subject of extensive investigation.
    [Show full text]